During fiscal year 2011, we accomplished the following: 1. Continued analysis of BLSA 2 year returnees using the scheme established for the first round of volunteers. Specifically, this involves analysis of protein and RNA in 4 samples per volunteer: 1) unactivated cells incubated at 37C, 2,3) cells activated with anti-CD3 for 2h and 4h, and 4) unactivated cells kept at 4C (to preserve in vivo signature of gene expression). 2. Completed analysis of preparative 2D protein expression assays in young and old. 3. Compared gene expression in CD4+T cells incubated at 37 C in normal medium (containing fetal calf serum) or defined synthetic medium (lacking fetal calf serum). We found that the NF-&#954;B-induced pathway was up-regulated and the BCR-activation pathway was down-regulated in the synthetic medium as previously noted in normal medium. One difference was that the NF-&#954;B-induced pathway was not at the top of the list of up-regulated pathways in the synthetic medium. Thus, NF&#954;B target genes are up-regulated by a cell-intrinsic mechanism upon metabolic activation by incubation at 37C. However, components in fetal calf serum may accentuate NF-&#954;B up-regulation in normal serum. 4. We initiated collaborative studies with Dr. Sam John and Gordon Hager (NCI) to examine the effects of anti-inflammatory corticosteroids on human B cells. Conditions for dexamethasone treatment and chromatin immunoprecipitation studies were first standardized using the Burkitt lymphoma cell line, BJAB. Additionally, we standardized procedures to purify nave and memory primary human B cells from apheresis packs.